Sulfonamide compound and crystal thereof

ABSTRACT

(S)-1-(4-Chloro-5-isoquinolinesulfonyl)-3-(methylamino)pyrrolidine monohydrochloride and a crystal thereof, and a crystal of the aforementioned monohydrochloride having a major peak or peaks at one or more positions selected from the group consisting of positions where 2θs are about 13.9°, 21.5°, 21.7°, 22.4°, 22.8°, 24.5° and 35.0° in a powder X-ray diffraction spectrum, which have excellent properties as active ingredient of a medicament for prophylactic and/or therapeutic treatment of glaucoma and the like.

TECHNICAL FIELD

The present invention relates to a novel sulfonamide compound and a saltthereof. More specifically, the present invention relates to(S)-1-(4-chloro-5-isoquinolinesulfonyl)-3-(methylamino)pyrrolidine,monohydrochloride thereof and monohydrobromide thereof useful as activeingredients of medicaments.

BACKGROUND ART

Sulfonamide derivatives which inhibit phosphorylation of myosinregulatory light chain and have an intraocular pressure reducing actionand neutrophil migration inhibitory action are known (InternationalPublication WO2007/026664), and it has been elucidated that thesesulfonamide derivatives are useful as active ingredients of medicamentsfor prophylactic and/or therapeutic treatment of glaucoma and the like.

However,(S)-1-(4-chloro-5-isoquinolinesulfonyl)-3-(methylamino)pyrrolidine infree base represented by the following formula (1) is not disclosed inInternational Publication WO2007/026664 (hereinafter in thespecification, this compound in a free base may be referred to as“Compound 1”).

Hydrochloride of Compound 1 mentioned above is disclosed inInternational Publication WO2007/026664, and this hydrochloride isprepared by treating tert-butoxycarbonylated Compound 1 with excessivehydrochloric acid and removing the tert-butoxycarbonyl group (Example19-3). However, the aforementioned publication discloses only thepreparation method, and no description is given as for number ofhydrochloric acid molecules added to the resulting(S)-1-(4-chloro-5-isoquinolinesulfonyl)-3-(methylamino)pyrrolidinehydrochloride, physicochemical properties of the resulting hydrochlorideand the like.

Patent document 1: WO2007/026664

DISCLOSURE OF THE INVENTION Object to be Achieved by the Invention

For use of(S)-1-(4-chloro-5-isoquinolinesulfonyl)-3-(methylamino)pyrrolidine as anactive ingredient of a medicament, an object of the present invention isto provide a novel salt form thereof having more preferred properties.

Means for Achieving the Object

The inventors of the present invention precisely conducted reproductiveexperiment of the method described in Example 19-3 of the aforementionedInternational Publication WO2007/026664 to prepare hydrochloride of(S)-1-(4-chloro-5-isoquinolinesulfonyl)-3-(methylamino)pyrrolidinedescribed in the above publication, and conducted researches on thestructure and physicochemical properties of the substance. As a result,they found that the hydrochloride was a salt having two hydrochloricacids added, i.e., (S)-1-(4-chloro-5-isoquinolinesulfonyl)3-(methylamino)pyrrolidine dihydrochloride (hereinafter in thespecification, this substance is sometimes referred to as“dihydrochloride”). They also found that changes in physicochemicalproperties of the dihydrochloride were observed after stability test at60° C. for two weeks, and that the hydrochloride gave remarkablehygroscopicity as well as the changes in physicochemical propertiesafter storage for two weeks under the conditions of 25° C. and 84% RH.

Generally as for substances as active ingredients of medicaments, it isknown that chemical or physical stability of the substancessignificantly influence on effectiveness and safety of the medicaments.Therefore, especially in industrial scale production, it is desirable touse a substance that is more stable against temperature or humidity asan active ingredient of a medicament. By using such a stable substance,reduction of content of an active ingredient during storage ordistribution of a medicament can be prevented, and thus a medicamentthat can ensure effectiveness and safety over a long period of time canbe supplied.

From the viewpoint as mentioned above for use of the above Compound 1 asa medicament, the inventors of the present invention conductedresearches in order to provide a substance in a form of a salt havingmore preferred properties, especially more improved properties instability and hygroscopicity than the aforementioned dihydrochloride. Asa result, it was found that both of(S)-1-(4-chloro-5-isoquinolinesulfonyl)-3-(methylamino)pyrrolidinemonohydrochloride (hereinafter in the specification, this substance issometimes referred to as “monohydrochloride”) and(S)-1-(4-chloro-5-isoquinolinesulfonyl)-3-(methylamino)pyrrolidinemonohydrobromide (hereinafter in the specification, this substance issometimes referred to as “monohydrobromide”) had good stability andreduced hygroscopicity. The present invention was accomplished on thebasis of the aforementioned findings.

The present invention thus provides the following substances.

-   (1)    (S)-1-(4-Chloro-5-isoquinolinesulfonyl)-3-(methylamino)pyrrolidine    monohydrochloride.-   (2)    (S)-1-(4-Chloro-5-isoquinolinesulfonyl)-3-(methylamino)pyrrolidine    monohydrobromide.-   (3) A crystal of    (S)-1-(4-chloro-5-isoquinolinesulfonyl)-3-(methylamino)pyrrolidine.-   (4) A crystal of    (S)-1-(4-chloro-5-isoquinolinesulfonyl)-3-(methylamino)pyrrolidine    monohydrochloride.-   (5) A crystal of    (S)-1-(4-chloro-5-isoquinolinesulfonyl)-3-(methylamino)pyrrolidine    monohydrobromide.-   (6) The crystal of    (S)-1-(4-chloro-5-isoquinolinesulfonyl)-3-(methylamino)pyrrolidine    according to (3), which has a major peak or peaks at one or more    positions selected from the group consisting of positions where 2θs    are about 9.1°, 13.8°, 21.0°, 21.7° and 23.6° in a powder X-ray    diffraction spectrum.-   (7) The crystal of    (S)-1-(4-chloro-5-isoquinolinesulfonyl)-3-(methylamino)pyrrolidine    according to (3) or (6), which has major peaks at position where 2θs    are about 9.1°, 13.8°, 21.0°, 21.7° and 23.6° in a powder X-ray    diffraction spectrum.-   (8) The crystal of    (S)-1-(4-chloro-5-isoquinolinesulfonyl)-3-(methylamino)pyrrolidine    according to (3), (6) or (7), which has major peaks at positions    where wave numbers are about 1335, 1146, 1139, 1096 and 609 cm⁻¹ in    an infrared absorption spectrum.-   (9) The crystal of    (S)-1-(4-chloro-5-isoquinolinesulfonyl)-3-(methylamino)pyrrolidine    according to (3), (6), (7) or (8), which has a fusion peak at about    106° C. in differential scanning calorimetry (temperature increasing    rate: 10° C./minute).-   (10) The crystal of    (S)-1-(4-chloro-5-isoquinolinesulfonyl)-3-(methylamino)pyrrolidine    monohydrochloride according to (4), which has a major peak or peaks    at one or more positions selected from the group consisting of    positions where 2θs are about 13.9°, 21.5°, 21.7°, 22.4°, 22.8°,    24.5° and 35.0° in a powder X-ray diffraction spectrum.-   (11) The crystal of    (S)-1-(4-chloro-5-isoquinolinesulfonyl)-3-(methylamino)pyrrolidine    monohydrochloride according to (4) or (10), which has major peaks at    positions where 2θs are about 13.9°, 21.5°, 21.7°, 22.4°, 22.8°,    24.5° and 35.0° in a powder X-ray diffraction spectrum.-   (12) The crystal of    (S)-1-(4-chloro-5-isoquinolinesulfonyl)-3-(methylamino)pyrrolidine    monohydrochloride according to (4), (10) or (11), which shows major    peaks at positions where wave numbers are about 1330, 1150, 1140 and    613 cm⁻¹ in an infrared absorption spectrum.-   (13) The crystal of    (S)-1-(4-chloro-5-isoquinolinesulfonyl)-3-(methylamino)pyrrolidine    monohydrochloride according to (4), (10), (11) or (12), which has a    decomposition peak at about 290% in differential scanning    calorimetry (temperature increasing rate: 10° C./minute).-   (14) The crystal of    (S)-1-(4-chloro-5-isoquinolinesulfonyl)-3-(methylamino)pyrrolidine    monohydrobromide according to (5), which has a major peak or peaks    at one or more positions selected from the group consisting of    positions where 2θs are about 21.3°, 22.4°, 24.1°, 30.7° and 34.8°    in a powder X-ray diffraction spectrum.-   (15) The crystal of    (S)-1-(4-chloro-5-isoquinolinesulfonyl)-3-(methylamino)pyrrolidine    monohydrobromide according to (5) or (14), which has major peaks at    positions where 2θs are about 21.3°, 22.4°, 24.1°, 30.7° and 34.8°    in a powder X-ray diffraction spectrum.-   (16) The crystal of    (S)-1-(4-chloro-5-isoquinolinesulfonyl)-3-(methylamino)pyrrolidine    monohydrobromide according to (5), (14) or (15), which has major    peaks at positions where wave numbers are about 2695, 1307, 1149,    1139 and 612 cm⁻¹ in an infrared absorption spectrum.-   (17) The crystal of    (S)-1-(4-chloro-5-isoquinolinesulfonyl)-3-(methylamino)pyrrolidine    monohydrobromide according to (5), (14), (15) or (16), which has a    decomposition peak at about 270° C. in differential scanning    calorimetry (temperature increasing rate: 10° C./minute).-   (18) A method for preparing the crystal of    (S)-1-(4-chloro-5-isoquinolinesulfonyl)-3-(methylamino)pyrrolidine    according to any one of (3) and (6) to (9), which comprises the    steps of adding a base to an acidic solution containing    (S)-1-(4-chloro-5-isoquinolinesulfonyl)-3-(methylamino)pyrrolidine    to neutralize the solution wherein the compound is prepared by    reacting    (S)-3-[N-(tert-butoxycarbonyl)-1-(4-chloro-5-isoquinolinesulfonyl)pyrrolidine    with an acid in a solvent, and isolating deposited solid.-   (19) A method for preparing the crystal of    (S)-1-(4-chloro-5-isoquinolinesulfonyl)-3-(methylamino)pyrrolidine    according to any one of (3) and (6) to (9), which comprises the    steps of adding a base to a solution of dihydrohalide of    (S)-1-(4-chloro-5-isoquinolinesulfonyl)-3-(methylamino)pyrrolidine    to neutralize the solution and thereby prepare    (S)-1-(4-chloro-5-isoquinolinesulfonyl)-3-(methylamino)pyrrolidine,    and isolating a solid of the compound deposited in a poor solvent in    which the compound is hardly dissolved.-   (20) A method for preparing the crystal of    (S)-1-(4-chloro-5-isoquinolinesulfonyl)-3-(methylamino)pyrrolidine    monohydrochloride according to any one of (4) and (10) to (13),    which comprises the steps of adding 0.5 to 2 equivalents of    hydrochloric acid to a solution in which    (S)-1-(4-chloro-5-isoquinolinesulfonyl)-3-(methylamino)pyrrolidine    is dissolved, and isolating a deposited crystal.-   (21) A method for preparing the crystal of    (S)-1-(4-chloro-5-isoquinolinesulfonyl)-3-(methylamino)pyrrolidine    monohydrobromide according to any one of (5) and (14) to (17), which    comprises the steps of adding 0.5 to 2 equivalents of hydrobromic    acid to a solution in which    (S)-1-(4-chloro-5-isoquinolinesulfonyl)-3-(methylamino)pyrrolidine    is dissolved, and isolating a deposited crystal.-   (22) A pharmaceutical composition comprising    (S)-1-(4-chloro-5-isoquinolinesulfonyl)-3-(methylamino)pyrrolidine    monohydrochloride or (S)-1-(4-chloro-5-isoquinolinesulfonyl)-3    (methylamino)pyrrolidine monohydrobromide as an active ingredient.-   (23) A pharmaceutical composition comprising the crystal according    to any one of (4) and (10) to (13) as an active ingredient.-   (24) A pharmaceutical composition comprising the crystal according    to any one of (5) and (14) to (17) as an active ingredient.-   (25) A composition comprising the monohydrochloride, wherein mass    ratio of the monohydrochloride is about 20% or more based on the    total mass of    (S)-1-(4-chloro-5-isoquinolinesulfonyl)-3-(methylamino)pyrrolidine,    salt thereof and solvate thereof, which is taken as 100%.

EFFECT OF THE INVENTION

(S)-1-(4-Chloro-5-isoquinolinesulfonyl)-3-(methylamino)pyrrolidinemonohydrochloride and(S)-1-(4-chloro-5-isoquinolinesulfonyl)-3-(methylamino)pyrrolidinemonohydrobromide provided by the present invention have a characteristicfeature that they are more stable and less hygroscopic compared with thedihydrochloride prepared by the method described in Example 19-3 ofInternational Publication WO2007/026664. Therefore, by using thesesubstances as active ingredients of medicaments, medicaments can beprovided in which reduction of a content of active ingredient duringstorage or distribution is suppressed, and medicaments can be stablysupplied of which effectiveness and safety can be ensured over a longperiod of time.

BRIEF DESCRIPTION OF DRAWINGS

FIG. 1 A figure depicting differential scanning calorimetry spectrum ofCompound 1 obtained by a method similar to the method described inExample 1

FIG. 2 A figure depicting differential scanning calorimetry spectrum ofthe monohydrochloride obtained by the method described in Example 3

FIG. 3 A figure depicting differential scanning calorimetry spectrum ofthe monohydrobromide obtained by the method described in Example 4

FIG. 4 A figure depicting powder X-ray diffraction spectrum of Compound1 obtained by the method described in Example 1

FIG. 5 A figure depicting powder X-ray diffraction spectrum of themonohydrochloride obtained by the method described in Example 3

FIG. 6 A figure depicting powder X-ray diffraction spectrum of themonohydrobromide obtained by the method described in Example 4

FIG. 7 A figure depicting powder X-ray diffraction spectrum of thedihydrochloride

FIG. 8 A figure depicting infrared absorption spectrum of Compound 1obtained by the method described in Example 1

FIG. 9 A figure depicting infrared absorption spectrum of themonohydrochloride obtained by the method described in Example 3

FIG. 10 A figure depicting infrared absorption spectrum of themonohydrobromide obtained by the method described in Example 4

FIG. 11 A figure depicting infrared absorption spectrum of thedihydrochloride

BEST MODE FOR CARRYING OUT THE INVENTION

This application is a patent application filed with claimingconventional priorities based on Japanese Patent Application No.2007-174323 filed in Japan on Jul. 2, 2007 and Provisional PatentApplication No. 61/034,222 filed in the United States on Mar. 6, 2008.The entire disclosures of the specifications, claims, and drawings ofthese applications are incorporated in the disclosure of the presentspecification by reference.

Compound 1 can be prepared by, for example, reacting(S)-3-[N-(tert-butoxycarbonyl)-1-(4-chloro-5-isoquinolinesulfonyl)pyrrolidineobtainable by the method described in WO2007/026664 with a large excessamount of an acid in a solvent to remove the tert-butoxycarbonyl group,then adding a base to the solution, and isolating deposited solid.

As the solvent used for the removal of the tert-butoxycarbonyl group,for example, water, alcohols such as methanol, ethanol and 2-propanol,ethers such as tetrahydrofuran and 1,4-dioxane, esters such as ethylacetate and isopropyl acetate, acetonitrile, dichloromethane and thelike are preferred, and they can be used as a mixture if needed. Amongthem, methanol, ethanol and 2-propanol are more preferred. Use of amixture of water and 2-propanol is particularly preferred. The mixingratio of water and 2-propanol is, for example, about 1:10 to 10:1, morepreferably 1:1 to 10:1, particularly preferably 2:1 to 6:1.

Type of the acid used for the removal of tert-butoxycarbonyl group isnot particularly limited, and ordinarily used mineral acids, organicacid and the like may be used. For example, hydrochloric acid, sulfuricacid, nitric acid, hydrobromic acid, phosphoric acid and trifluoroaceticacid are preferred, hydrochloric acid and trifluoroacetic acid are morepreferred, and hydrochloric acid is particularly preferred.

Amount of the acid used for the removal of tert-butoxycarbonyl group isnot particularly limited, and the acid may be added to such an extentthat the removal reaction sufficiently proceeds. The amount may be, forexample, 2 equivalents or more, more preferably 2.0 to 10.0 equivalents,particularly preferably 2.0 to 5.0 equivalents, based on Compound 1.

As the reaction temperature, a suitable temperature of, for example,from 1° C. to the reflux temperature of the solvent may be chosen, and apreferred example is a temperature of from 30 to 70° C. The reactiontime is, for example, usually about 0.1 to 24 hours, more preferably 0.5to 10 hours, particularly preferably 1 to 5 hours. Progress of thereaction can be monitored by thin layer chromatography (TLC), highperformance liquid chromatography (HPLC) or the like, and an acidicsolution of Compound 1 can be prepared usually by appropriatelyterminating the reaction when the yield of Compound 1 reaches to themaximum.

Although type of the base added to the acidic solution of Compound 1 todeposit Compound 1 produced is not particularly limited, for example, aninorganic base is preferred. Examples of the inorganic base include, forexample, alkali metal bases such as sodium hydroxide, potassiumhydroxide, sodium carbonate, potassium carbonate, sodiumhydrogencarbonate, sodium methoxide and potassium t-butoxide and thelike, sodium hydroxide, potassium hydroxide and the like are morepreferred, and sodium hydroxide is particularly preferred. These bases,per se, can be used as solid, or they can also be used by beingdissolved beforehand in water or an alcohol such as methanol, ethanol,or 2-propanol. By preparing an aqueous solution containing a base at agiven concentration beforehand and using the solution, an advantage isenjoyed that an amount of the base to be added may be easily adjusted.

As the crystallization solvent used for depositing Compound 1, forexample, water, alcohols such as methanol, ethanol and 2-propanol,ethers such as tetrahydrofuran, esters such as ethyl acetate andisopropyl acetate, acetonitrile, dichloromethane and the like arepreferred, and a mixture of these solvents can be used if needed. Amongthem, water, methanol, ethanol, and 2-propanol are more preferred.Further, use of a mixture of water and 2-propanol is particularlypreferred. An example of a mixing ratio of water and 2-propanolincludes, for example, about 1:10 to 10:1. A ratio of 1:1 to 10:1 ismore preferred, and 2:1 to 6:1 is particularly preferred. When a solventdifferent from the reaction solvent used for the removal oftert-butoxycarbonyl group is used as a solvent for the crystallization,the solvent can be replaced by concentration or the like.

Although an amount of the base to be added is not particularly limitedand the base may be added in an amount providing a good yield ofCompound 1 as a solid. Generally, an example includes about 1 equivalentor more based on 1 equivalent of the acid added. It is also possible toselect the amount of the base to be added depending on a pH of thesolution. Generally, it is preferable to adjust a pH of the solution tobe 7 or higher, and more preferred example includes a pH of from 8 to12.

Temperature of the solution at the time of adding the base is notparticularly limited so far that the temperature is an appropriatetemperature of from 0° C. to the boiling point of the solution. Atemperature in the range of 10 to 40° C. is more preferred.

Although deposition concentration of Compound 1 after the addition ofthe base may vary depending on the type of a solvent used or dependingon a mixing ratio of solvents in a case of a mixed solvent, a lowerlimit includes, for example, generally 1 w/v % or higher, preferably 5w/v % or higher. As an upper limit, 30 w/v % or lower is preferred, andmore preferred example includes 15 w/v % or lower. For example, when amixed solvent of water and 2-propanol is used as the solvent, it ispreferred that the ratio thereof is 4:1 to 6:1 and the depositionconcentration is from 5 w/v % to 10 w/v %, and more preferred exampleincludes about 8 w/v %.

For deposition of a solid of Compound 1, another preferred embodimentincludes the addition of a small amount of Compound 1 as seed crystalsto a solution after the addition of a base.

Examples of the method for isolating Compound 1 deposited include knownmethods such as filtration and decantation. It is usually preferable toconduct isolation by filtration. Although the isolation of Compound 1 byfiltration may also be performed immediately after the addition of abase, the isolation is preferably performed after the deposition ofsolid reaches to a stationary state. For example, the isolation ispreferably performed 1 hour after the addition of the base, morepreferably performed 3 hours after the addition of the base.

At the time of isolating Compound 1 deposited, it is also possible tocool the solution after the addition of the base and then perform theisolation. Examples of the method for cooling include a method ofcooling rapidly, a method of cooling step by step, a method of coolinggradually over a period of time, a method of standing the solution forcooling and the like, and the method of cooling step by step, the methodof cooling gradually over a period of time, and the method of standingthe solution for cooling are more preferred. Generally, a coolingtemperature is preferably from 0 to 20° C., and more preferably from 0to 10° C.

Compound 1 isolated can be dried by a drying method ordinarilyperformed, for example, drying under reduced pressure, drying by warmingunder reduced pressure, drying by warming with ventilation, air dryingand the like. The drying by warming under reduced pressure and thedrying by warming with ventilation are particularly preferred. Whenwarming is applied for the drying, a temperature of room temperature orhigher is usually chosen. A temperature of from 40 to 60° C. is morepreferred. As for time period for the drying, drying may be performeduntil residual level of a solvent becomes appropriate. For example, 10hours or longer is preferred.

Among the preparation methods described above, a preferred embodimentincludes the following method.

An acidic solution of Compound 1 is prepared by adding(S)-3-[N-(tert-butoxycarbonyl)-1-(4-chloro-5-isoquinolinesulfonyl)pyrrolidineand hydrochloric acid in an amount of 3.0 equivalents based on thecompound to a mixed solvent of water and 2-propanol at a mixing ratio of2:1 to 6:1 and performing a reaction at a temperature of 50 to 65° C.for 1 to 3 hours with stirring. The acidic solution is added with sodiumhydroxide at a temperature of 20 to 35° C. with stirring to adjust a pHat from 8 to 10, then the solution is further stirred for 1 to 20 hours,and deposited solid is isolated. By drying the isolated solid underreduced pressure at 50° C. for 10 hours or longer, crystals of Compound1 are obtained.

Compound 1 can also be isolated by dissociating the added hydrochloricacids from the dihydrochloride obtained according to the methoddescribed in Example 19-3 of WO2007/026664 by a reaction with a base inan appropriate solvent to prepare a solution containing Compound 1, thenremoving the solvent by concentration, and adding a poor solvent inwhich Compound 1 is hardly dissolved to the residue to deposit Compound1.

As the reaction solvent used for the dissociation of the hydrochloricacid from the dihydrochloride, for example, water, alcohols such asmethanol, ethanol and 2-propanol, ethers such as tetrahydrofuran and1,4-dioxane, esters such as ethyl acetate and isopropyl acetate,acetonitrile, dichloromethane and the like are preferred, and a mixtureof these solvents may be used if needed. Among them, water, methanol,ethanol and 2-propanol are more preferred, and water is particularlypreferred.

As the base added for a purpose of dissociating the hydrochloric acidfrom the dihydrochloride, the base added to the acidic solution ofCompound 1 can be used. Generally, an amount of the base to be added ispreferably 1.6 equivalents or more, more preferably 2 to 4 equivalents,based on 1 equivalent of the dihydrochloride. A temperature of thesolution at the time of adding the base is not particularly limited sofar that the temperature is an appropriate temperature of from 0° C. tothe boiling point of the solution. A range of 5 to 25° C. is morepreferred. As for the method of adding the base, the base can usually beadded at one portion with stirring the solution. The base may be addedseveral times as divided portions or continuously added over a period oftime by a method such as dropping or the like.

For the removal of a solvent by concentration, it is also possible toconduct substitution with a solvent having a lower boiling pointbeforehand such as by extraction and then perform the concentration. Apreferred example includes, for example, a method of extracting Compound1 with an organic solvent such as dichloromethane from the aqueoussolution containing Compound 1 prepared by the aforementioned method andevaporating dichloromethane under reduced pressure.

Examples of the poor solvent (i.e., a solvent in which Compound 1 ishardly dissolved) added to deposit Compound 1 from the residue includewater, ethyl acetate, n-hexane, n-heptane, diisopropyl ether and thelike, and ethyl acetate and n-hexane are more preferred. These solventscan also be used as a mixture if needed.

Compound 1 deposited can be isolated by the method explained above, andcan be dried if needed.

The structure of Compound 1 can be confirmed on the basis of ¹H-¹Hcorrelation, ¹³C-¹³C correlation, ¹H-¹³C correlation and the like in anuclear magnetic resonance spectrum and/or analysis of mass spectrum.For example, the structure can be confirmed on the basis of ¹H-¹Hcorrelation in a nuclear magnetic resonance spectrum, and the (m/Z)value of the protonated compound (326) in a mass spectrum.

The monohydrochloride can be prepared by adding hydrochloric acid to asolution in which Compound 1 is dissolved, and isolating depositedcrystals. Compound 1 may be in the form of either crystal or amorphous,or a mixture thereof. As the solvent for dissolving Compound 1, alcoholssuch as methanol, ethanol, 1 propanol and 2-propanol, ethers such astetrahydrofuran and 1,4-dioxane, esters such as ethyl acetate andisopropyl acetate, acetone, acetonitrile and the like are preferred, andthese solvents can be used as a mixture if needed. Among them, methanol,ethanol, 1-propanol and 2-propanol are more preferred, and ethanol and2-propanol are particularly preferred. These solvents may contain waterat a ratio of about 30% or less in terms of volume ratio.

An amount of the solvent added for dissolving Compound 1 by using thesolvents as mentioned above may vary depending on the type of thesolvent used and a mixing ratio in case of a mixed solvent. An amountwherein Compound 1 is dissolved at a temperature below the boiling pointof the solvent used is preferred, and further from a viewpoint of anyield of crystals obtained, use of an amount wherein Compound 1 isdissolved and to give a saturation concentration at a temperature nearthe boiling point of the solvent is particularly preferred. Morespecifically, when 2-propanol is used as the solvent, for example, amethod of adding 100 to 200 ml of 2-propanol to 10 g of Compound 1 andwarming the mixture to 60° C. or higher is preferred, and when ethanolis used, a preferred example includes a method of adding 70 to 150 ml ofethanol to 10 g of Compound 1 and warming the mixture to 60° C. orhigher. When insoluble solids exist, it is preferred to remove theinsoluble solids from the solution by an operation such as filtration orthe like.

As for an amount of hydrochloric acid added to the aforementionedsolution to deposit the crystals of the monohydrochloride, a range offrom 0.5 to 2 equivalents is generally preferred based on Compound 1. Arange of from 0.8 to 1.5 equivalents is more preferred, 0.9 to 1.2equivalents is particularly preferred. A most particularly preferredexample includes 0.95 to 1.05 equivalents. When a solvent is chosen fromwhich the monohydrochloride is preferentially deposited, it is alsopossible to add 2 equivalents or more of hydrochloric acid. For example,when ethanol or 2-propanol is chosen as the solvent, a preferred exampleincludes a range of from 0.5 to 10 equivalents as the amount ofhydrochloric acid, and a more preferred example includes a range of 0.5to 5 equivalents. Hydrochloric acid to be added can be used after beingdissolved in water or the aforementioned solvents. A use of an aqueoushydrochloric acid solution prepared beforehand at a given concentrationis preferred from a viewpoint of convenience of controlling the amountto be added.

Although a temperature at the time of adding the hydrochloric acid isnot particularly limited so far that the temperature is an appropriatetemperature of from 0° C. to the boiling point of the solution, atemperature that gives saturated concentration of Compound 1 or higheris preferred. Specifically, when 10 g of the compound is dissolved in100 to 200 ml of 2-propanol, for example, it is preferable to add 5 Naqueous hydrochloric acid at 40 to 60° C.

A method for adding hydrochloric acid is not particularly limited.Generally, the acid may be added as one portion with stirring thesolution, or the acid may be added several times as divided portions, orcontinuously added over a period of time by a method such as dropping orthe like.

For deposition of crystals, preferred embodiments include a method ofadding a small amount of crystals of the monohydrochloride as seedcrystals to the solution after addition of hydrochloric acid, and amethod of cooling the solution after addition of hydrochloric acid.Examples of the method for cooling include a method of cooling rapidly,a method of cooling step by step, a method of cooling gradually over aperiod of time, a method of standing the solution for cooling and thelike. More preferred examples include the method of cooling step bystep, the method of cooling gradually over a period of time, and themethod of standing the solution for cooling.

A final concentration of the monohydrochloride at the time of thedeposition of crystals after the addition of hydrochloric acid may varydepending on type of the solvent used and a mixing ratio in the case ofa mixed solvent. An example of a lower limit generally includes 0.5 w/v% or higher, more preferable example includes 1 w/v % or higher. Apreferred example of an upper limit includes 20 w/v % or lower, and morepreferred example includes 10 w/v % or lower. Specifically, for example,when a mixed solvent of water and 2-propanol (mixing ratio is 1:9 to0.5:9.5) is used as the solvent for deposition, a final concentration of2.5 to 10 w/v % is preferred, and more preferred example includes 5 to7.5 w/v %.

Examples of the method for isolating deposited crystals include knownmethods such as filtration and decantation. Generally, isolation byfiltration is preferred. Although the isolation of the crystals may beperformed immediately after the addition of hydrochloric acid, theisolation is preferably performed after the deposition of crystalsreaches to a stationary state. For example, the isolation is preferablyperformed 1 hour after the addition, and more preferably performed 3hours after the addition.

For collection of the deposited crystals, collection of the crystalsafter cooling of the solution in which deposition of crystals reaches toa stationary state is preferred from a viewpoint of an yield of thecrystals to be obtained and the like. Examples of the method for coolinginclude a method of cooling rapidly, a method of cooling step by step, amethod of cooling gradually over a period of time, a method of standingthe solution for cooling and the like. The method of cooling step bystep, the method of cooling gradually over a period of time, and themethod of standing the solution for cooling are more preferred. As acooling temperature, a temperature of from 0 to 20° C. is generallypreferred, and 0 to 10° C. is more preferred.

After the crystals are isolated by filtration, the crystals can bewashed with the solvent used for dissolving Compound 1, for example,ethanol, 2-propanol or a mixed solvent of water therewith, which is aneffective operation for removing impurities. Examples of the method forwashing include a method of rinsing the crystals on a filter with asolvent, and a method of putting the crystals into a solvent to form asuspension, sufficiently stirring the suspension, and then collectingthe crystals again by filtration. Furthermore, it is also effective toperform both of the aforementioned two kinds of washing operations.

The collected crystals can be dried by a drying method ordinarilyperformed, for example, drying under reduced pressure, drying by warmingunder reduced pressure, drying by warming with ventilation, air dryingand the like.

Among the aforementioned preparation methods, a preferred exampleincludes a method of warming a suspension of Compound 1 in 2-propanol to50 to 60° C. to dissolve Compound 1, adding dropwise 1 equivalent ofhydrochloric acid based on Compound 1 to the solution at 20 to 60° C.with stirring, and further stirring for 1 to 20 hours to obtaincrystals.

The monohydrobromide can be prepared by adding hydrobromic acid to asolution in which Compound 1 is dissolved, and isolating depositedcrystals. This method can be performed in the same manner as that of theaforementioned method for preparing the monohydrochloride by usinghydrobromic acid instead of hydrochloric acid.

For evaluating the type of the acid forming a salt with Compound 1 andthe number of the acid added, ion exchange chromatography is applied tocalculate the number of the acid added per Compound 1. For example, amethod comprises calculation of the number of the added acid perCompound 1 by dissociating the added acid by ion exchange using an ionexchange column such as DIONEX IonPacAS14 with an internal diameter of 4mm and a length of 25 cm, and comparing peak areas with those ofstandard solutions of known ion concentrations by using an electricconductivity detector to quantify the acid.

Further, the type of the acid forming a salt with Compound 1 and thenumber of the acid added can also be evaluated by other means such asquantification of amounts of elements by elemental analysis.Furthermore, if the substance consists of a single kind of crystal, thetype of the acid forming a salt with Compound 1 and the number of acidadded can also be evaluated by X-ray structure analysis.

It is well-known to those skilled in the art that the number of addedacid measured by ion chromatography may include some measurement errorsdue to various kinds of factors. As for the number of the added acid perCompound 1, a measurement error of ±0.2 is usually acceptable, and ameasurement error of ±0.1 is acceptable for a more preferredmeasurement.

As a confirmation test of the monohydrochloride or the monohydrobromide(hereinafter these substances are also referred to as “substance in theform of a salt”), powder X-ray diffractometry may also be used.Furthermore, an infrared absorption spectrum may be measured. Morespecifically, an example of the method includes a method of measuring aninfrared absorption spectrum by using powder. For example, the potassiumbromide tablet method described in Japanese Pharmacopoeia, General TestMethods, “Infrared Absorption Spectrometry” can be chosen.

For evaluation of purity of Compound 1 or the substance in the form of asalt, the area percentage method based on HPLC is convenient. Forevaluation of water content of Compound 1 or the substance in the formof a salt, the volumetric titration method, the coulometric titrationmethod described in Japanese Pharmacopoeia, General Test Methods, “WaterDetermination”, the loss on drying test and the like can be used. Whensample weight is small, the coulometric titration method can bepreferably chosen.

When it is necessary to measure an amount of Compound 1 or the substancein the form of a salt contained in a pharmaceutical preparation, use ofHPLC is usually convenient and thus preferred. For example, acalibration curve can be prepared by HPLC using standard samples ofCompound 1 having known chemical purities, and amount of Compound 1existing in a sample can be determined on the basis of the calibrationcurve.

Examples of an optical system used for the powder X-ray diffractionspectrometry include an ordinary light concentrating type optical systemand a parallel beam type optical system. Although the optical system tobe used is not particularly limited, when resolution and intensity aredesired to be ensured, the measurement is preferably performed by usinga light concentrating type optical system. Further, when orientation isdesired to be suppressed, which is a phenomenon that molecules arearranged along a certain direction depending on shapes of crystals(needle shape, tabular shape and the like), the measurement ispreferably performed by using a parallel beam type optical system.Examples of measurement apparatus of the light concentrating typeoptical system include XRD-6000 (Shimadzu), MultiFlex (RigakuInternational) and the like. Examples of measurement apparatus of theparallel beam type optical system include XRD-7700 (Shimadzu), RINT2200Ultima+/PC (Rigaku International) and the like.

It is well-known to those skilled in the art that 2θ values in a powderX-ray diffraction spectrum may include some measurement errors due tovarious kinds of factors. A measurement error of usually about ±0.3°,typically about ±0.2°, or about ±0.1 for a more preferable measurement,is acceptable. Therefore, it will be understood by those skilled in theart that values indicated in the specification with the term “about” for2θ values may include an acceptable measurement error.

Although it is well-known to those skilled in the art that a measuredvalue obtained by differential scanning calorimetry is a numerical valuecharacteristic to crystals as a measurement object, it is alsowell-known to those skilled in the art that, besides measurement errors,melting point may occasionally change in a practical measurement due tovarious causes such as contamination of an acceptable amount ofimpurities. Therefore, those skilled in the art can understand that thepeak temperatures mentioned in the specification practically measured indifferential scanning calorimetry may occasionally change, and that adegree of the change may be, for example, about ±5° C., typically about±3° C., or about ±2° C. for preferred measurement. Examples ofmeasurement apparatus used for the differential scanning calorimetryinclude PYRIS Diamond DSC (Perkin-Elmer), DSC 3200 (Bruker AXS) and thelike.

Some measurement error is accepted also for the wave number in infraredabsorption spectrum, and those skilled in the art readily understandthat it is acceptable that values described in the specification mayinclude such measurement errors. For example, according to the 4thedition of European Pharmacopoeia, in comparison with a referencespectrum in a confirmation test based on infrared absorption spectra,coincidence of wave number scale with a difference of ±0.5% or less isaccepted. Although it is not intended to be bound by the aforementionedcriterion in the specification, for example as an example of criterion,a measurement error of about ±0.8%, preferably about ±0.5%, particularlypreferably about ±0.2%, is acceptable for the wave number scale.

Thermal stability of Compound 1 or the substance in the form of a saltcan be evaluated by, for example, sealing a sample in a glass vial orthe like, storing the vial under a severe temperature condition such as40 to 80° C. in a dark place for a given period of time, and thenobserving or measuring appearance, purity, water content and the like ofCompound 1 or the substance in the form of a salt. Change of puritybefore and after storage especially serves as an important index ofthermal stability. For example, the evaluation is preferably performedunder a storage condition of 60° C.

Hygroscopicity of Compound 1 or the substance in the form of a salt canbe evaluated by putting a sample on a glass weighing dish, storing thedish in a dark place for a given period of time in an open state at atemperature of 25 to 40° C. under a humidification condition of about 75to 94% relative humidity, and then observing or measuring appearance,purity, water content and the like of Compound 1 or the substance in theform of a salt. Increase of water content before and after storageespecially serves as an important hygroscopic index. For example, theevaluation is preferably performed after storage conditions of 25° C.and 84% RH.

If a composition containing the monohydrochloride satisfies theconditions; 1) the ratio of the monohydrochloride exceeds 0%, and 2) anyeffect of the monohydrochloride is observed when compared with acomposition that is equivalent except for the content of themonohydrochloride being 0%, it should be understood that any of suchcomposition falls within the scope of the present invention. Further, acomposition in which even a trace amount of the monohydrochloride isdetected falls within the scope of the present invention.

In a composition containing the monohydrochloride, when(S)-1-(4-chloro-5-isoquinolinesulfonyl)-3-(methylamino)pyrrolidine, saltthereof and solvate thereof are focused and a total mass thereof isconsidered to be 100%, a mass ratio of the monohydrochloride isgenerally preferred to be about 90% or more, more preferably near 100%.

From a viewpoint of control of hygroscopicity of a composition that alsocontains the dihydrochloride, when at least 20% of the monohydrochlorideis contained, the hygroscopicity controlling effect can be observed.Therefore, the ratio of at least 20% or more is exemplified as apreferred embodiment.

Further, from a viewpoint of suppression of coloring of the compositionthat also contains the dihydrochloride with passage of time, themonohydrochloride is preferably contained in an amount of about 60% ormore, and the content of about 80% or more is exemplified as a morepreferred embodiment.

The same shall apply to a composition containing the monohydrobromide.

EXAMPLES

The present invention will be more specifically explained with referenceto examples. However, the scope of the present invention is not limitedby the following examples.

Example 1(S)-1-(4-Chloro-5-isoquinolinesulfonyl)-3-(methylamino)pyrrolidine

(S)-3-[N-(tert-Butoxycarbonyl)-1-(4-chloro-5-isoquinolinesulfonyl)pyrrolidine(370 g) obtained by the method described in WO2007/026664 was suspendedin a mixed solution of 2-propanol (740 ml) and water (1261 ml). Thissuspension was added with hydrochloric acid (35%, specific gravity:1.18, 271 g, Kanamori Industry) at 23° C. with stirring, and the mixturewas warmed to 59.5° C. to react for 2 hours with stirring. Aftercompletion of the reaction, the temperature of the reaction mixture wasmaintained at 26 to 28° C., adjusted to pH 8.47 by adding dropwise 2mol/l aqueous sodium hydroxide (1350 ml) with stirring, and then addedwith (S)-1-(4-chloro-5-isoquinolinesulfonyl)-3-(methylamino)pyrrolidine(1.42 g) as seed crystals at 26.0° C. Further, the reaction mixture wasleft to cool to 18.0° C. over 20 hours and 40 minutes with stirring.After the stand for cooling, the reaction mixture gave pH of 8.18. Themixture was adjusted to pH 9.67 by adding 2 mol/l aqueous sodiumhydroxide (150 ml). After stirring for 1 hour, the reaction mixture wascooled to 1.0° C. over 4 hours and 22 minutes, and the deposited solidwas collected by suction filtration using a Buchner funnel (internaldiameter: 240 mm, filter paper: No. 131). The pale brown wet solidobtained was dried under reduced pressure at 50° C. for 18 hours toobtain slightly brown crystals of the title compound (259.3 g).

Example 2(S)-1-(4-Chloro-5-isoquinolinesulfonyl)-3-(methylamino)pyrrolidine

(S)-1-(4-Chloro-5-isoquinolinesulfonyl)-3-(methylamino)pyrrolidinedihydrochloride (1.50 g) obtained by the method described inWO2007/026664 was dissolved in water (32 ml). This solution wasvigorously stirred, and slowly added dropwise with 2 N aqueous sodiumhydroxide (4.13 ml, Wako Pure Chemical Industries) under ice cooling.The resulting suspension was further stirred at room temperature for 1hour, and added with dichloromethane (30 ml), and the organic layer wasseparated. The aqueous layer was extracted with dichloromethane (30 ml),and the combined organic layers were washed with water (50 ml) and driedover anhydrous magnesium sulfate. The solvent was evaporated underreduced pressure, and ethyl acetate (10 ml) and n-hexane (20 ml) wereadded to the residue. The deposited solid was collected by filtrationand dried by warming at 50° C. for 20 hours under reduced pressure toobtain the title compound (1.07 g).

Example 3(S)-1-(4-Chloro-5-isoquinolinesulfonyl)-3-(methylamino)pyrrolidinemonohydrochloride

(S)-1-(4-Chloro-5-isoquinolinesulfonyl)-3-(methylamino)pyrrolidine (150g) obtained in Example 1 was suspended in 2-propanol (2400 ml), and thesuspension was warmed to 61.5° C. to dissolve the compound. Thissolution was filtered by using a membrane filter (internal diameter: 90mm, ADVANTEC PTFE, 0.2 μm), and the filtrate was added dropwise with 300ml of a mixed solution of hydrochloric acid (35%, specific gravity:1.18, 48.0 g, Kanamori Industry) and purified water (261.4 ml, FukujuPharmaceuticals) with maintaining the filtrate at 55° C. The mixture wasstirred at 55° C. for 44 minutes, then cooled to 2.0° C. over 2 hoursand 11 minutes, and further stirred for 1 hour and 28 minutes, and thenthe deposited crystals were separated by suction filtration using aBuchner funnel (internal diameter: 150 mm, filter paper: No. 5C). Thepale brown wet crystals obtained were dried under reduced pressure at50° C. for 14 hours and 35 minutes to obtain the title compound (152 g).

Example 4(S)-1-(4-Chloro-5-isoquinolinesulfonyl)-3-(methylamino)pyrrolidinemonohydrobromide

(S)-1-(4-Chloro-5-isoquinolinesulfonyl)-3-(methylamino)pyrrolidine (100mg) obtained in Example 1 was dissolved in ethanol (2.5 ml), and addedwith 48% hydrobromic acid (33.2 μL, Wako Pure Chemical Industries), andthe mixture was stirred at room temperature for 17 hours. The reactionmixture was further added with ethanol (4 ml), and the deposited solidwas collected by filtration and dried by warming at 60° C. for 24 hoursunder reduced pressure to obtain the title compound (93.5 mg).

Test Example 1 Nuclear Magnetic Resonance Spectrum (¹H-NMR)

The compound obtained by the method described in Example 1 was taken inan amount of 0.02 g, and dissolved in 0.6 ml of dimethyl sulfoxide-d₆(deuterated solvent) containing 0.05% of tetramethylsilane as aninternal reference substance, and the nuclear magnetic resonancespectrum was measured under the following conditions. The substance gavepeaks at δ (ppm): 1.95 (1H, m), 2.13-2.19 (2H, m), 2.33 (3H, s),3.31-3.33 (2H, m), 3.49-3.53 (3H, m), 7.84-7.88 (1H, dd), 8.48-8.50 (1H,d), 8.63-8.65 (1H, d), 8.76 (1H, s), 9.41 (1H, s), and the ¹H-¹Hcorrelation of these peaks supported the structure of Compound 1.

The compound obtained by the method described in Example 3 was taken inan amount of 0.02 g, and dissolved in 0.6 ml of heavy water (deuteratedsolvent) containing 0.05% of sodium 3-trimethylsilylpropionate-d₄ as aninternal reference substance, and the nuclear magnetic resonancespectrum was measured under the following conditions. The substance gavepeaks at δ (ppm): 2.40-2.45 (1H, m), 2.70-2.75 (1H, m), 2.90 (3H, s),3.65-3.71 (1H, m), 3.78-3.85 (2H, m), 3.96-4.01 (1H, m), 4.16-4.19 (1H,m), 4.33 (3H, s), 7.62-7.66 (1H, dd), 8.05-8.09 (2H, dd), 8.40 (1H, s),8.83 (1H, s), and the ¹H-¹H correlation of these peaks supported thestructure of the monohydrochloride.

Conditions:

Nuclear magnetic resonance apparatus: JNM LA400 (JEOL)Oscillation frequency: 400 MHz

Nuclide: ¹H Test Example 2 Mass Spectrum

Mass spectrum of the compound obtained by the method described inExample 1 was measured under the following conditions, and a protonatedmolecule was detected at (m/z)=326, which result supported the structureof Compound 1.

Mass spectrum of the compound obtained by the method described inExample 3 was measured under the following conditions, and a protonatedmolecule was detected at (m/z)=326, which result supported the structureof the monohydrochloride.

Conditions:

Mass spectrometer: JMS-SX102 (JEOL)Ionizing method: FAB

Detected ion: Cation

Dissolution solvent: Dimethyl sulfoxideMatrix: m-Nitrobenzyl alcohol

Test Example 3 Differential Scanning Calorimetry

Differential scanning calorimetry of the compound obtained by a methodsimilar to the method described in Example 1 was performed under thefollowing conditions, and the spectrum shown in FIG. 1 was obtained, inwhich a thermal absorption peak considered to be a fusion peak wasobserved at 107° C.

Conditions: Calorimeter: PYRIS Diamond DSC

Temperature increasing condition: Increase from 50° C. to 250° C. at arate of 10° C./minute

Differential scanning calorimetry of the compound obtained by the methoddescribed in Example 3 was performed under the following conditions, andthe spectrum shown in FIG. 2 was obtained, in which a thermal absorptionpeak considered to be a resolution peak was observed at 290° C.

Conditions: Calorimeter: PYRIS Diamond DSC

Temperature increasing condition: Increase from 50° C. to 350° C. at arate of 10° C./minute

Differential scanning calorimetry of the compound obtained by the methoddescribed in Example 4 was performed under the following conditions, andthe spectrum shown in FIG. 3 was obtained, in which a thermal absorptionpeak considered to be a resolution peak was observed at 270° C.

Conditions Calorimeter: PYRIS Diamond DSC

Temperature increasing condition: Increase from 50° C. to 350° C. at arate of 10° C./minute

Test Example 4 Ion Exchange Chromatography

When the compound obtained by the method described in Example 1 wasanalyzed by ion exchange chromatography under the following conditions,fluoride ion, chloride ion, bromide ion, nitrite ion, nitrate ion,phosphate ion, sulfate ion and other anions were not detected, and thusit was confirmed that the compound obtained by the method described inExample 1 was(S)-1-(4-chloro-5-isoquinolinesulfonyl)-3-(methylamino)pyrrolidine(Compound 1).

When salt number of the compound obtained by the method described inExample 3 was confirmed by ion exchange chromatography under thefollowing conditions, 1.0 of chloride ion was observed per molecule of(S)-1-(4-chloro-5-isoquinolinesulfonyl)-3-(methylamino)pyrrolidine, andthus it was confirmed that the substance was a salt of(S)-1-(4-chloro-5-isoquinolinesulfonyl)-3-(methylamino)pyrrolidine(Compound 1) added with one hydrochloric acid (monohydrochloride).

When salt number of the compound obtained by the method described inExample 4 was confirmed by ion exchange chromatography under thefollowing conditions, 1.0 of bromide ion was observed per molecule of(S)-1-(4-chloro-5-isoquinolinesulfonyl)-3-(methylamino)pyrrolidine, andthus it was confirmed that the substance was a salt of(S)-1-(4-chloro-5-isoquinolinesulfonyl)-3-(methylamino)pyrrolidine(Compound 1) added with one hydrobromic acid (monohydrobromide).

Conditions:

Sample concentration: 100 μg/mlIon chromatograph: DIONEX ICS-1000 (Dionex Japan)Detector: Electric conductivity detectorColumn: DIONEX IonPacAS14, internal diameter: 4 mm, length: 25 cmGuard column: DIONEX IonPacAG14, internal diameter: 4 mm, length: 5 cmColumn temperature: 30° C.Mobile phase: 1.0 mmol/l Aqueous sodium hydrogencarbonate containing 3.5mmol/l sodium carbonateFlow rate: About 1.2 ml/minute

Injection: 10 μl

Suppressor: ASRS-ULTRA (recycling mode, SRS 24 mA)

Test Example 5 Powder X-Ray Diffractometry

Powder X-ray diffractometry of the compound obtained by the methoddescribed in Example 1 was performed under the following conditions, andthe diffraction spectrum shown in FIG. 4 was obtained. In this powderX-ray diffraction spectrum, characteristic major peaks were observed atthe positions where 2θs are 9.1°, 13.8°, 21.0°, 21.7° and 23.6°. Peakswere also observed at the positions of 17.4°, 20.5°, 26.3°, 28.0° and30.2°, and any one or more of these peaks are also considered to be apeak or peaks characteristic to Compound 1. Furthermore, peaks were alsoobserved at the positions of 24.1°, 25.7°, 28.7°, 32.7°, 38.3° and42.0°, and any one or more of these peaks can also be considered to be apeak or peaks characteristic to Compound 1.

Powder X-ray diffractometry of the monohydrochloride obtained by themethod described in Example 3 was performed under the followingconditions, and the diffraction spectrum shown in FIG. 5 was obtained.In this powder X-ray diffraction spectrum, characteristic major peakswere observed at the positions where 2θs are 13.9°, 21.5°, 21.7°, 22.4°,22.8°, 24.5° and 35.0°. Peaks were also observed at the positions of14.3°, 28.2°, 29.3°, 30.8° and 36.0°, and any one or more of these peakscan also be considered to be a peak or peaks characteristic to themonohydrochloride. Furthermore, peaks were also observed at thepositions of 8.9°, 18.4°, 36.4° and 39.1°, and any one or more of thesepeaks can also be considered to be a peak or peaks characteristic to themonohydrochloride.

Powder X-ray diffractometry of the monohydrobromide obtained by themethod described in Example 4 was performed under the followingconditions, and the diffraction spectrum shown in FIG. 6 was obtained.In this powder X-ray diffraction spectrum, characteristic major peakswere observed at the positions where 2θs are 21.3°, 22.4°, 24.1°, 30.7°and 34.8°. Peaks were also observed at the positions of 13.8°, 21.7°,27.3°, 28.6° and 29.1°, and any one or more of these peaks can also beconsidered to be a peak or peaks characteristic to the monohydrobromide.Furthermore, peaks were also observed at the positions of 16.8°, 19.4°,25.6°, 27.1° and 36.0°, and any one or more of these peaks can also beconsidered to be a peak or peaks characteristic to the monohydrobromide.

Compound 1, the monohydrochloride, and the monohydrobromide were judgedto be crystals by visual inspection, and they were further confirmed tobe crystals by the aforementioned powder X-ray diffraction analysis. Itwas also confirmed that Compound 1, the monohydrochloride, and themonohydrobromide gave powder X-ray diffraction spectra different fromthat of the dihydrochloride shown in FIG. 7.

Measurement Conditions:

X-Ray diffractometer: XRD-6000 (Shimadzu) or RINT 2200 Ultima+/PC(Rigaku International)X-Ray source: CuKα (40 kV, 30 mA)Operation mode: ContinuousScanning speed: 2°/minuteScanning axis: θ to 2θScanning range: 5 to 600Scattering slit: 1°Light-receiving slit: 0.30 mm

Test Example 6 Infrared Absorption Spectrum

Infrared absorption spectrum of Compound 1 obtained by the methoddescribed in Example 1 was measured under the following conditions, andthe spectrum shown in FIG. 8 was obtained. In this infrared absorptionspectrum, characteristic absorptions were observed at the positionswhere wave numbers are 1335, 1146, 1139, 1096 and 609 cm⁻¹. Absorptionswere also observed at the positions of 1219, 1156, 1130, 1033, 1027,1000, 766, 742 and 584 cm⁻¹, and any one or more of these absorptionscan be considered to be a characteristic absorption or absorptions ofCompound 1.

Infrared absorption spectrum of the monohydrochloride obtained by themethod described in Example 3 was measured under the followingconditions, and the spectrum shown in FIG. 9 was obtained. In thisinfrared absorption spectrum, characteristic absorptions were observedat the positions where wave numbers are 1330, 1150, 1140 and 613 cm⁻¹.Absorptions were also observed at the positions of 2747, 2695, 2690,1487, 1091 and 1046 cm⁻¹, and any one or more of these absorptions canbe considered to be a characteristic absorption or absorptions of themonohydrochloride.

Infrared absorption spectrum of the monohydrobromide obtained by themethod described in Example 4 was measured under the followingconditions, and the spectrum shown in FIG. 10 was obtained. In thisinfrared absorption spectrum, characteristic absorptions were observedat the positions where wave numbers are 2695, 1307, 1149, 1139 and 612cm⁻¹. Absorptions were also observed at the positions of 2963, 2932,2916, 2909, 2880, 2807, 2795, 2751, 1466, 1222, 1219, 1089 and 1044cm⁻¹, and any one or more of these absorptions can be considered to be acharacteristic absorption or absorptions of the monohydrobromide.

It was also confirmed that Compound 1, the monohydrochloride, and themonohydrobromide gave spectra different from the infrared absorptionspectrum of the dihydrochloride shown in FIG. 11.

Measurement Conditions:

Infrared spectrophotometer: FTIR-8300 (Shimadzu)Measurement method: Potassium bromide tablet methodControl: Potassium bromide tablet

Gain: Auto Aperture: Auto

Minimum wave number: 400 cm⁻¹Maximum wave number: 4000 cm⁻¹Number of integration: 45 times

Detector: Standard

Apodization function: Happ-GenzelDecomposition: 2 cm⁻¹Mirror velocity: 2.8

Test Example 7 Purity Test

Purity of Compound 1 obtained by the method described in Example 1 wasmeasured by high performance liquid chromatography under the followingconditions. As a result, a peak of(S)-1-(4-chloro-5-isoquinolinesulfonyl)-3-(methylamino)pyrrolidine wasobserved at from 12 to 13 minutes, and the purity was found to be 99.9%.

Purity of the monohydrochloride obtained by the method described inExample 3 was similarly measured. As a result, a peakof(S)-1-(4-chloro-5-isoquinolinesulfonyl)-3-(methylamino)pyrrolidine wasobserved at from 12 to 13 minutes, and the purity was found to be 99.4%.

Purity of the monohydrobromide obtained by the method described inExample 4 was similarly measured. As a result, a peakof(S)-1-(4-chloro-5-isoquinolinesulfonyl)-3-(methylamino)pyrrolidine wasobserved at from 12 to 13 minutes, and the purity was found to be 99.7%.

Conditions of high performance liquid chromatography:High performance liquid chromatography apparatus: LC-10A Series(Shimadzu) or Agilent 1100 series (Agilent Technologies)Solution concentration: 500 μg/ml

Injection: 10 μl

Detector: Ultraviolet absorptiometer (measurement wavelength: 245 nm)Column: XBridge Shield RP18 5 μm, internal diameter: 4.6 mm, length: 15cm (Waters)Column temperature: Constant temperature around 40° C.Mobile phase A: 20 mmol/l Sodium phosphate buffer (pH 7.0)Mobile phase B: AcetonitrileLiquid feeding program: A concentration gradient was formed by changingthe mixing ratio of the mobile phase A and the mobile phase B as shownin Table 1.Flow rate: 1.0 ml/minute

TABLE 1 Liquid feeding program Time from injection (minute) Mobile phaseA (%) Mobile phase B (%)   0 to 45.5 80 → 15 20 → 85 45.5 to 54   15 8554 to 69 80 20Dissolution solvent: Mixture of water/methanol (1:1)

Test Example 8 Measurement of Water Content

Water content of the monohydrochloride obtained by the method describedin Example 3 and the monohydrobromide obtained by the method describedin Example 4 was measured according to the “coulometric titrationmethod” defined in Japanese Pharmacopoeia, General Test Methods, “WaterDetermination” under the following conditions, and water contents of themonohydrochloride was found to be 0.20% and that of the monohydrobromidewas found to be 0.14%.

Conditions:

Water content measurement apparatus: AQ-7 (Hiranuma Sangyo)Sample amount: 5 mg

Anolyte: Aqualyte RS (Sigma-Aldrich) Catholyte: Aqualyte CN(Sigma-Aldrich) Test Example 9 Thermal Stability Test

The monohydrochloride obtained by the method described in Example 3, themonohydrobromide obtained by the method described in Example 4, and thedihydrochloride as a control were weighed in a glass vial in an amountof 40 mg each, and the glass vial was sealed. Each sample was stored ina dark place at 60° C. for 2 to 4 weeks, and after the storage,appearance was evaluated by visual inspection. Further, purity wasmeasured under the conditions of Test Example 7, and water content wasmeasured under the conditions of Test Example 8.

The dihydrochloride gave change of appearance after the storage at 60°C. for 2 weeks. Whilst, the monohydrochloride and the monohydrobromidegave no change of appearance and no degradation of purity even after thestorage at 60° C. for 4 weeks.

TABLE 2 Before After 2 After 4 Salt Test storage weeks weeksMonohydrochloride Appearance White White White powder powder powderWater content 0.20% 0.06% 0.08% Purity 99.4% 99.9% 99.7%Monohydrobromide Appearance White White White powder powder powder Watercontent 0.14% 0.05% 0.05% Purity 99.7% 99.9% 99.8% DihydrochlorideAppearance White Pale brown —* powder powder Water content 3.76% 2.93%—* Purity 98.1% 99.5% —* *The dihydrochloride gave apparent change ofappearance after 2 weeks, and therefore the stability test wasterminated.

Test Example 10 Hygroscopicity Test 1

The monohydrochloride obtained by the method described in Example 3, themonohydrobromide obtained by the method described in Example 4, and thedihydrochloride as a control were weighed on a glass weighing dish in anamount of 40 mg each, and each sample was stored in a dark place at 25°C. and 84% RH for 2 to 4 weeks. After the storage, appearance wasevaluated by visual inspection. Further, purity was measured under theconditions of Test Example 7, and water content was measured under theconditions of Test Example 8.

The dihydrochloride gave apparent change of appearance and markedmoisture absorption after the storage at 25° C. and 84% RH for 2 weeks.Whilst, the monohydrochloride and the monohydrobromide gave no change ofappearance as well as no substantial increase of water content and nodegradation of purity even after the storage at 25° C. and 84% RH for 4weeks.

It was confirmed by Test Examples 9 and 10 that the monohydrochlorideand the monohydrobromide have more satisfactory thermal stability andremarkably lower hygroscopicity compared with the dihydrochloride.

TABLE 3 Before After 2 After 4 Salt Test storage weeks weeksMonohydrochloride Appearance White White White powder powder powderWater content 0.20% 0.23% 0.26% Purity 99.4% 99.8% 99.6%Monohydrobromide Appearance White White White powder powder powder Watercontent 0.14% 0.05% 0.05% Purity 99.7% 99.9% 99.7% DihydrochlorideAppearance White Brown —** powder powder Water content 3.76% 15.24%  —**Purity 98.1% 99.3% —** **The dihydrochloride showed definite change ofappearance and marked moisture absorption after 2 weeks, and thereforethe stability test was terminated.

Test Example 11 Hygroscopicity Test 2

Compound 1 obtained by a method similar to the method described inExample 1 (two kinds of lots), the monohydrochloride obtained by themethod described in Example 3, the dihydrochloride, and mixtures of themonohydrochloride and the dihydrochloride at ratios of 9:1, 8:2, 6:4,4:6 and 2:8 were weighed on a glass weighing dish in an amount of each40 mg, and each sample was stored in a dark place at 25° C. and 84% RHfor 2 to 4 weeks. After the storage, appearance was evaluated by visualinspection. Further, purity was measured under the conditions of TestExample 7, and water content was measured under the conditions of TestExample 8.

Both of the two lots of Compound 1 gave moisture absorption after thestorage at 25° C. and 84% RH for 2 weeks, and further moistureabsorption after the storage at 25° C. and 84% RH for 4 weeks.

The mixtures of the monohydrochloride and the dihydrochloride containingthe dihydrochloride at a ratio of 40% or more gave apparent change ofappearance after the storage at 25° C. and 84% RH for 2 weeks. It wasalso observed that a higher ratio of the dihydrochloride gave higherhygroscopicity.

It was confirmed by Test Example 11 that the monohydrochloride had ahygroscopicity lower than any of Compound 1 (free base) and mixtures ofthe monohydrochloride and the dihydrochloride. Moreover, it was alsoconfirmed that, in a mixture of the dihydrochloride and themonohydrochloride, increase of the mixing ratio of the monohydrochloridelowered the hygroscopicity of the mixture.

TABLE 4 Before After 2 After 4 Salt Test storage weeks weeks Compound 1Appearance Slightly Slightly Slightly (Lot 1) brownish brownish brownishwhite white white powder powder powder Water  0.9%  5.7%  8.1% contentPurity  99.9%  99.9% 99.9% Compound 1 Appearance Slightly SlightlySlightly (Lot 2) brownish brownish brownish white white white powderpowder powder Water  1.7%  7.6%  9.1% content Purity  99.9%  99.9% 99.9%Monohydrochloride Appearance White White —*** powder powder Water  0.0% 0.1% —*** content Purity 100.0% 100.0% —*** Dihydrochloride AppearanceWhite Pale —*** powder yellow powder Water  5.2%  19.8% —*** contentPurity 100.0%  99.9% —*** Monohydrochloride:Dihydrochloride AppearanceWhite White —*** Mixing ratio 9:1 powder powder Water  0.7%  2.1% —***content Purity 100.0% 100.0% —*** Monohydrochloride:dihydrochlorideAppearance White White —*** Mixing ratio 8:2 powder powder Water  0.9% 6.6% —*** content Purity 100.0% 100.0% —***Monohydrochloride:dihydrochloride Appearance White Slightly —*** Mixingratio 6:4 powder yellowish white powder Water  1.7%  11.2% —*** contentPurity 100.0%  99.9% —*** Monohydrochloride:dihydrochloride AppearanceWhite Pale —*** Mixing ratio 4:6 powder yellow powder Water  4.3%  16.8%—*** content Purity 100.0%  99.9% —*** Monohydrochloride:dihydrochlorideAppearance White Pale —*** Mixing ratio 2:8 powder yellow powder Water 4.3%  18.4% —*** content Purity 100.0%  99.9% —*** ***Test was notperformed.

INDUSTRIAL APPLICABILITY

(S)-1-(4-Chloro-5-isoquinolinesulfonyl)-3-(methylamino)pyrrolidinemonohydrochloride and(S)-1-(4-chloro-5-isoquinolinesulfonyl)-3-(methylamino)pyrrolidinemonohydrobromide provided by the present invention are characterized inthat they are highly stable and have low hygroscopicity. Therefore,these substances are useful as active ingredients of medicaments, ofwhich decrease in content of the active ingredient during storage ordistribution is suppressed, and also useful for stable supply ofmedicaments for which efficacy and safety can be ensured over a longperiod of time.

1. (S)-1-(4-Chloro-5-isoquinolinesulfonyl)-3-(methylamino)pyrrolidine monohydrochloride.
 2. (S)-1-(4-Chloro-5-isoquinolinesulfonyl)-3-(methylamino)pyrrolidine monohydrobromide.
 3. A crystal of (S)-1-(4-chloro-5-isoquinolinesulfonyl)-3-(methylamino)pyrrolidine.
 4. A crystal of (S)-1-(4-chloro-5-isoquinolinesulfonyl)-3-(methylamino)pyrrolidine monohydrochloride.
 5. A crystal of (S)-1-(4-chloro-5-isoquinolinesulfonyl)-3-(methylamino)pyrrolidine monohydrobromide.
 6. The crystal of (S)-1-(4-chloro-5-isoquinolinesulfonyl)-3-(methylamino)pyrrolidine according to claim 3, which has a major peak or peaks at one or more positions selected from the group consisting of positions where 2θs are about 9.1°, 13.8°, 21.0°, 21.7° and 23.6° in a powder X-ray diffraction spectrum.
 7. The crystal of (S)-1-(4-chloro-5-isoquinolinesulfonyl)-3-(methylamino)pyrrolidine according to claim 3 or 6, which has major peaks at positions where 2θs are about 9.1°, 13.8°, 21.0°, 21.7° and 23.6° in a powder X-ray diffraction spectrum.
 8. The crystal of (S)-1-(4-chloro-5-isoquinolinesulfonyl)-3-(methylamino)pyrrolidine according to claim 3, 6 or 7, which has major peaks at positions where wave numbers are about 1335, 1146, 1139, 1096 and 609 cm⁻¹ in an infrared absorption spectrum.
 9. The crystal of (S)-1-(4-chloro-5-isoquinolinesulfonyl)-3-(methylamino)pyrrolidine according to claim 3, 6, 7 or 8, which has a fusion peak at about 106° C. in differential scanning calorimetry (temperature increasing rate: 10° C./minute).
 10. The crystal of (S)-1-(4-chloro-5-isoquinolinesulfonyl)-3-(methylamino)pyrrolidine monohydrochloride according to claim 4, which has a major peak or peaks at one or more positions selected from the group consisting of the positions where 2θs are about 13.9°, 21.5°, 21.7°, 22.4°, 22.8°, 24.5° and 35.0° in a powder X-ray diffraction spectrum.
 11. The crystal of (S)-1-(4-chloro-5-isoquinolinesulfonyl)-3-(methylamino)pyrrolidine monohydrochloride according to claim 4 or 10, which has major peaks at positions where 2θs are about 13.9°, 21.5°, 21.7°, 22.4°, 22.8°, 24.5° and 35.0° in a powder X-ray diffraction spectrum.
 12. The crystal of (S)-1-(4-chloro-5-isoquinolinesulfonyl)-3-(methylamino)pyrrolidine monohydrochloride according to claim 4, 10 or 11, which has major peaks at positions where wave numbers are about 1330, 1150, 1140 and 613 cm⁻¹ in an infrared absorption spectrum.
 13. The crystal of (S)-1-(4-chloro-5-isoquinolinesulfonyl)-3-(methylamino)pyrrolidine monohydrochloride according to claim 4, 10, 11 or 12, which has a decomposition peak at about 290° C. in differential scanning calorimetry (temperature increasing rate: 10° C./minute).
 14. The crystal of (S)-1-(4-chloro-5-isoquinolinesulfonyl)-3-(methylamino)pyrrolidine monohydrobromide according to claim 5, which has a major peak or peaks at one or more positions selected from the group consisting of positions where 2θs are about 21.3°, 22.4°, 24.1°, 30.7° and 34.8° in a powder X-ray diffraction spectrum.
 15. The crystal of (S)-1-(4-chloro-5-isoquinolinesulfonyl)-3-(methylamino)pyrrolidine monohydrobromide according to claim 5 or 14, which has major peaks at positions where 2θs are about 21.3°, 22.4°, 24.1°, 30.7° and 34.8° in a powder X-ray diffraction spectrum.
 16. The crystal of (S)-1-(4-chloro-5-isoquinolinesulfonyl)-3-(methylamino)pyrrolidine monohydrobromide according to claim 5, 14 or 15, which has major peaks at positions where wave numbers are about 2695, 1307, 1149, 1139 and 612 cm⁻¹ in an infrared absorption spectrum.
 17. The crystal of (S)-1-(4-chloro-5-isoquinolinesulfonyl)-3-(methylamino)pyrrolidine monohydrobromide according to claim 5, 14, 15 or 16, which shows a decomposition peak at about 270° C. in differential scanning calorimetry (temperature increasing rate: 10° C./minute).
 18. A method for preparing the crystal of (S)-1-(4-chloro-5-isoquinolinesulfonyl)-3-(methylamino)pyrrolidine according to any one of claims 3 and 6 to 9, which comprises the steps of adding a base to an acidic solution containing (S)-1-(4-chloro-5-isoquinolinesulfonyl)-3-(methylamino)pyrrolidine to neutralize the solution, wherein said compound is prepared by reacting (S)-3-[N-(tert-butoxycarbonyl)-1-(4-chloro-5-isoquinolinesulfonyl)pyrrolidine with an acid in a solvent, and isolating a deposited solid.
 19. A method for preparing the crystal of (S)-1-(4-chloro-5-isoquinolinesulfonyl)-3-(methylamino)pyrrolidine according to any one of claims 3 and 6 to 9, which comprises the steps of adding a base to a solution of dihydrohalide of (S)-1-(4-chloro-5-isoquinolinesulfonyl)-3-(methylamino)pyrrolidine to neutralize the solution and thereby prepare (S)-1-(4-chloro-5-isoquinolinesulfonyl)-3-(methylamino)pyrrolidine, and isolating a solid of the compound deposited in a poor solvent in which the compound is hardly dissolved.
 20. A method for preparing the crystal of (S)-1-(4-chloro-5-isoquinolinesulfonyl)-3-(methylamino)pyrrolidine monohydrochloride according to any one of claims 4 and 10 to 13, which comprises the steps of adding 0.5 to 2 equivalents of hydrochloric acid to a solution in which (S)-1-(4-chloro-5-isoquinolinesulfonyl)-3-(methylamino)pyrrolidine is dissolved, and isolating a deposited crystal.
 21. A method for preparing the crystal of (S)-1-(4-chloro-5-isoquinolinesulfonyl)-3-(methylamino)pyrrolidine monohydrobromide according to any one of claims 5 and 14 to 17, which comprises the steps of adding 0.5 to 2 equivalents of hydrobromic acid to a solution in which (S)-1-(4-chloro-5-isoquinolinesulfonyl)-3-(methylamino)pyrrolidine is dissolved, and isolating a deposited crystal.
 22. A pharmaceutical composition comprising (S)-1-(4-chloro-5-isoquinolinesulfonyl)-3-(methylamino)pyrrolidine monohydrochloride or (S)-1-(4-chloro-5-isoquinolinesulfonyl)-3-(methylamino)pyrrolidine monohydrobromide as an active ingredient.
 23. A pharmaceutical composition comprising the crystal according to any one of claims 4 and 10 to 13 as an active ingredient.
 24. A pharmaceutical composition comprising the crystal according to any one of claims 5 and 14 to 17 as an active ingredient. 